OPERATING INSTRUCTIONS AND SYSTEM DESCRIPTION FOR
Contents
1. D Electronic Instruments for the Life Sciences pease ade OPERATING INSTRUCTIONS AND SYSTEM DESCRIPTION FOR THE ELP 01D ELECTROPORATOR FOR TISSUE AND CULTURED CELLS ELP 01D VERSION 1 1 npi 2014 npi electronic GmbH Bauhofring 16 D 71732 Tamm Germany Phone 49 0 7141 9730230 Fax 49 0 7141 9730240 support npielectronic com http www npielectronic com ELP 01D User Manual Table of Contents 17 Safety IR CCU AL ONS nseni in tases sate aatas tog itd E sen Sacia tid E idea ted tard dasa 3 2 Special Safety Notice for High Voltage Instrument ssseeeseseesseseeeseeeresseseresresseseresresse 4 ie EBP OTD ceca tlc A a ea eect e a e a a dete 5 Dy E aR DIE 011S n AE EE EEE TA A ET 5 3 2 Descrpton ot the Front Pan l ereersse e A E E ES N 5 4 Operatheater ge EE E slow E ES EEE SR 8 5 Literature with Focus on Methods s sesssseesssseessesssesssesesseessseesseesseesseessseeesseeesseesseesse 9 R SCH CALs ENA 22 59 iss E ana capes E E E A E E anes 9 version 1 1 page 2 ELP 01D User Manual 1 Safety Regulations VERY IMPORTANT Instruments and components supplied by npi electronic are NOT intended for clinical use or medical purposes e g for diagnosis or treatment of humans or for any other life supporting system npi electronic disclaims any warranties for such purpose Equipment supplied by npi electronic must be operated only by selected trained and adequately instructed perso2. or other components from the instrument or 19 cabinet TEMPERATURE DRIFT WARM UP TIME All analog electronic systems are sensitive to temperature changes Therefore all electronic instruments containing analog circuits should be used only in a warmed up condition i e after internal temperature has reached steady state values In most cases a warm up period of 20 30 minutes is sufficient HANDLING Please protect the device from moisture heat radiation and corrosive chemicals version 1 1 page 3 ELP 01D User Manual 2 Special Safety Notice for High Voltage Instruments 1 2 3 4 5 6 HIGH VOLTAGE RISK OF ELECTROCUTION Observe extreme caution when working with this instrument Always connect high voltage power supplies to protective earth Do not touch connections unless the instrument is turned off and the capacitance of both the load and power supply are earthed Allow adequate time for discharge of internal capacitance of the power supply Do not ground yourself or work under wet or damp conditions Servicing should be only done by qualified personnel aware of the hazards If in doubt return to supplier for servicing version 1 1 page 4 ELP 01D User Manual 3 ELP 01D 3 1 System Description The ELP 01D was designed as an easy to use device for electroporating tissue and cultured cells using square wave voltage pulses of up to 110 V The ELP 01D has two digital
3. For instance if you preset 10 cycles and you press start or a TTL pulse is applied the timer is reset to zero 10 cycles will be executed and the ELP stops still showing 10 at the display Upon the next start signal via switch or TTL the timer is reset again and 10 cycles will be executed 8 TRIGGER IN TTL connector BNC connector for connecting an external TRIGGER device minimum length of the TTL signal 10 us 9 START switch Switch for starting the timer momentary switch to upper position If the switch is in lower position permanent the timer can be started using an external TTL TRIGGER at 8 version 1 1 page 6 ELP 01D User Manual 10 REPETITIVE CONTINUOUS switch Switch for setting the operation mode of the ELP 01D REPETITIVE The timer cycles repetitively pulses The number cycles is of set by CYCLES switch 5 CONTINUOUS The timer cycles pulses continuously as long as the RESET STOP switch is used 11 Chassis connector Connector for grounding the cabinet 12 ISOLATED OUTPUT plugs The ISOLATED OUTPUT signal is available at two plugs red and blue The polarity of the 0 switch 14 is not changed i e the red plug is positive if the output polarity switch is set to or the analog input signal is positive This signal is completely isolated from earth Caution These plugs can provide HIGH VOLTAGE 13 INPUT TTL connector BNC connector for an external gating signal
4. If the internal timing unit should not be used a TTL signal from an external gating unit can be connected to 13 for generating a PULSE with an AMPLITUDE set by 2 The PULSE width is determined by the length of the TTL signal 14 0 switch Switch to set the polarity of the PULSE for electroporation In 0 position the OUTPUT is disabled 15 TEST ON button Push button for the electrode resistance test The resistance of the electrode is shown at 1 as long as the button is pressed version 1 1 page 7 ELP 01D User Manual 4 Operation The ELP 01D can be operated either by a gate applied to BNC connector 13 see above or by using the internal timing unit When using the timing unit the ELP 01D can be started either manually by pushing START 9 Figure 1 or by a TTL HI signal rising edge at TRIGGER TTL 8 Figure 1 This signal must be at least 10 us The time course of the timed ISOLATED OUTPUT is illustrated in Figure 2 50V p DELAY PULSE 1 DELAY PULSE 2 signal at ISOLATED OUTPUT if AMPLITUDE a is set top 50V time start 5V signal at TRIGGER TTL time Figure 2 scheme of timed operation After the DELAY the PULSE starts immediately After the end of the PULSE the second cycle begins with the DELAY and so on i e the time lag between the pulses is determined by the DELAY x 0 1 ms switch 3 Figure 1 For additional control a BNC connector labeled CURRENT O
5. UTPUT 0 1V mA is provided at the rear panel This BNC connector monitors the current that flows during a PULSE The OUTPUT is not isolated version 1 1 page 8 ELP 01D User Manual 5 Literature with Focus on Methods Ll Boudes M Pieraut S Valmier J Carroll P Scamps F Single cell electroporation of adult sensory neurons for gene screening with RNA interference mechanism J Neurosci Methods 2008 May 30 170 2 204 11 L Gal JS Morozov YM Ayoub AE Chatterjee M Rakic P Haydar TF Molecular and morphological heterogeneity of neural precursors in the mouse neocortical proliferative zones J Neurosci 2006 Jan 18 26 3 1045 56 L Martinez C Y P J Hollenbeck 2003 Transfection of primary CNS and PNS neurons by electroporation Methods in Cell Biology 71 321 332 Murphy RC Messer A Gene transfer methods for CNS organotypic cultures a comparison of three nonviral methods Mol Ther 2001 Jan 3 1 113 21 L Rathenberg J Nevian T Witzemann V High efficiency transfection of individual neurons using modified electrophysiology techniques J Neurosci Methods 2003 Jun 15 126 1 91 8 L Saito T Nakatsuji N Efficient gene transfer into the embryonic mouse brain using in vivo electroporation Dev Biol 2001 Dec 1 240 1 237 46 _ Tabata H Nakajima K Efficient in utero gene transfer system to the developing mouse brain using electroporation visualization of neuronal migration in the developing cortex N
6. euroscience 2001 103 4 865 72 6 Technical Data ELP 01D PULSE x 0 1 ms switch 5 digits thumb wheel 0 1 ms to 9999 9 ms digital DELAY x 0 1 ms switch 5 digits thumb wheel 0 1 ms to 9999 9 ms digital PULSE AMPLITUDE x1V switch 3 digits thumb wheel 0 V to 110 V digital TRIGGER IN TTL signal 10 us minimum ISOLATED OUTPUT 110 V max isolated set in 1 V steps CURRENT OUTPUT BNC connector 0 1 V mA Power Supply external 13 8 V DC version 1 1 page 9
7. nnel For details please consult the GENERAL TERMS OF DELIVERY AND CONDITIONS OF BUSINESS of npi electronic D 71732 Tamm Germany 1 2 3 4 5 GENERAL This system is designed for use in scientific laboratories and must be operated by trained staff only General safety regulations for operating electrical devices should be followed AC MAINS CONNECTION While working with the npi systems always adhere to the appropriate safety measures for handling electronic devices Before using any device please read manuals and instructions carefully The device is to be operated only at 115 230 Volt 60 50 Hz AC Please check for appropriate line voltage before connecting any system to mains Always use a three wire line cord and a mains power plug with a protection contact connected to ground protective earth Before opening the cabinet unplug the instrument Unplug the instrument when replacing the fuse or changing line voltage Replace fuse only with an appropriate specified type STATIC ELECTRICITY Electronic equipment is sensitive to static discharges Some devices such as sensor inputs are equipped with very sensitive FET amplifiers which can be damaged by electrostatic charge and must therefore be handled with care Electrostatic discharge can be avoided by touching a grounded metal surface when changing or adjusting sensors Always turn power off when adding or removing modules connecting or disconnecting sensors headstages
8. switches for setting PULSE duration and DELAY time The ELP 01D can be used continuously or for a selectable number of cycles A switch allows manual start and stop while cycling but the timing unit can also be operated using an external TRIGGER ELP 01D can also be used independently from the internal timing unit using a TTL INPUT for gating the PULSE The AMPLITUDE is set by a digital switch in 1 V steps An electrode resistance test unit allows to measure the electrode resistance up to 10 kQ 3 2 Description of the Front Panel npi RESSTANCE x0 N AMPLITUDE x1 V CYCLES CYCLE C bk DELAY pO Ima PULSE pO Irra nooo SOLATED OUTPUT CAUTION HGH VOLUIAGE AOOAO 9 Figure 1 ELP 01D front panel view 1 RESISTANCE x0 01 kQ display LED indicating the resistance of the electrode when operating the electrodes resistance test using button 15 2 AMPLITUDE x1V switch 3 digit thumb wheel switch to set the AMPLITUDE of the electroporation pulse range 0 110 V resolution 1 V The polarity is set by switch 14 version 1 1 page 5 ELP 01D User Manual 3 DELAY x0 1 ms switch 5 digit thumb wheel switch to set a delay time minimum delay time 100 us maximum delay time 9999 9 ms The delay time is the time from the rising edge of the trigger TTL signal to the beginning of the rising edge of the PULSE see also Figure 2 Important A DELAY of 0 is not allowed and leads to an undefined sta
9. te of the electroporator 4 PULSE x0 1 ms switch 5 digit thumb wheel switch to set the duration width of the PULSE minimum pulse width 100 us maximum pulse width 9999 9 ms 5 CYCLES switch 3 digit thumb wheel switch for setting the number of desired cycles 1 to 999 Note At the start of each train of cycles a RESET is executed i e the ELP will be reset immediately after a start signal comes either via manual switch or via TTL signal For instance if you preset 10 cycles and you press start or a TTL pulse is applied the timer is reset to zero 10 cycles will be executed and the ELP stops still showing 10 at the display Upon the next start signal via switch or TTL the timer is reset again and 10 cycles will be executed Important RESET has to be pressed after setting a new number of cycles 6 CYCLE COUNTER display LED indicating the number of cycles which had been carried out since the first start In CONTINUOUS mode the LED counts continuously 7 RESET STOP switch Switch for stopping the timer RESET the timer is stopped and the CYCLE COUNTER is set to zero STOP the timer is stopped and the CYCLE COUNTER shows the number of cycles which had been carried out If started again with switch 9 the CYCLE COUNTER continues to count Note At the start of each train of cycles a RESET is executed i e the ELP will be reset immediately after a start signal comes either via manual switch or via TTL signal
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